INFRAFRONTIER axenic (germ-free) service

INFRAFRONTIER2020 project and microbiome research

One of the INFRAFRONTIER2020 project aims is to develop pilot platforms and services supporting microbiome research. A service to derive germ-free (axenic) mice is now provided by three infrastructures with a long standing expertise and track record in the derivation of axenic and gnotobiotic mice, namely the Gnoto / Axenic Facility of the Instituto Gulbenkian de Ciência, the Karolinska Institutet Core Facility for Germ-free Research, and the CNRS Service Isotechnie of the PHENOMIN-TAAM infrastructure. The Karolinska Institutet has maintained a unique platform for the handling and derivation of gnotobiotic models kept under germ-free (GF) conditions since the 1960s. All infrastructures are partners and founding members of the ECGnoto network.

The participating INFRAFRONTIER infrastructures are among very few dedicated facilities in Europe who have the required equipment and expertise to generate germ-free and gnotobiotic mice. All infrastructures routinely offer their services to external users. The participating axenic- and gnotobiology platforms support research into host-microbiota interactions to study the role of the microbiome in both health and disease. This has been the subject of extensive research and established the involvement of the microbiome in metabolism, nutrition, physiology, and immune function and showed that mammalian microbiota play crucial roles in the pathogenesis of many diseases.

Access to the axenic service will be granted on the basis of scientific excellence and supports pilot projects for the derivation of-germ-free mice. Further breeding and characterisation of axenic mice or the development of gnotobiotic models can be offered on a fee-for-service or on a collaborative basis.

During the course of the INFRAFRONTIER2020 project a total of 2 Trans-national Access calls for the derivation of germ-free mice supporting a total of 10 projects will be published. The second call will be released in spring 2018.

The EC Horizon 2020 funded INFRAFRONTIER2020 project (2017 – 2020) supports eligible customers with a free of charge mouse axenic service implemented as a Trans-national Access activity providing a total of 5 access units in its first call.

The access unit offered covers the derivation of germ-free (axenic) mice from a breeding nucleus or from frozen materials provided by the applicants. Mice will be kept under germ-free conditions for 6-8 weeks.

Mutant mouse lines will be provided by the selected applicants as live animals (minimum of 10 females and 5 males) or quality controlled frozen embryos or sperm (minimum of 40 embryos or 2 straws of sperm). The participating infrastructures will transfer and breed mice under SPF conditions, and then introduce and foster 2 litters or 8 animals into one germ-free isolator until 2 months of age. Tissues or body fluids can be provided or arrangements are made for live germ-free shipments.

Costs: The access to the INFRAFRONTIER2020 axenic service is free of charge. However, the shipment cost of the starting material (breeding pairs or frozen material) to the infrastructures as well as a possible live germ-free animal shipment must be covered by the applicants. Extended breeding and other complementary services can be offered on a fee-for service or on a collaborative basis.

Eligibility: The INFRAFRONTIER2020 Trans-national Access call is open and proposals can be submitted from applicants around the world. Four projects must be allocated to applicants from EU Member States and Associated Countries, and one project can be allocated to applicants from third countries.

Application: Service requests for the INFRAFRONTIER2020 mouse axenic service can be made via this application form. Applications for the Trans-national Access activity must include a short description of the project involving the germ-free mouse mutant being generated by the INFRAFRONTIER2020 TA service.

Selection procedure: Proposals from eligible customers for free of charge access to the INFRAFRONTIER2020 mouse axenic service will be subject to a review procedure. The review will be based on short descriptions of the projects involving the mouse mutants that will be made germ-free by the TA service. A mixed panel of members of INFRAFRONTIER and of an external Evaluation Committee will assess service requests supported by the TA activity. In addition to scientific merit of applicants, soundness of the proposal, relevance for microbiome research and feasibility will be assessed. Applicants will be informed on the outcome of the evaluation within 6 weeks after the end of the call for which the TA application was submitted. All applications will be handled with strict confidentiality.

Acknowledgements: Please do acknowledge any support under this scheme in all resulting publications with "Part of this work has been funded by the European Union Research and Innovation programme Horizon 2020 (Grant Agreement Number 730879). The participating infrastructure which provided the service should be specifically mentioned in any publication resulting from the service.

Disclaimer: Certain mouse lines may not breed under axenic conditions, and derivation attempts will then be discontinued. Two attempts will be made to derive germ-free mice for each access unit.

Support offered

The participating axenic / gnotobiology platforms provide technological support to perform state of the art experiments relying on the removal and / or manipulation of mouse microbiota in a safe and controlled manner. In addition, they support researchers with access to organs, tissues and biofluids from germ-free mice, and provide a fast access to mice that are kept under germ-free conditions.

The services are provided to institutional, national and international users and the demand is growing due to the renewed interest in microbiome research.

Beyond the Trans-national Access service provision selected applicants will be further assisted with logistics support and advice to ship rederived germ-free mice to their facilities. Dedicated shippers will be provided for transportation and specialised courier services will be recommended.

The funded access unit covers the derivation of a single mouse mutant line into a germ-free condition. The simultaneous derivation of any additional line(s) can be negotiated with the participating infrastructure. Furthermore, extended breeding services under germ-free conditions can be offered either on a fee-for-service basis or on a collaborative basis for joint research projects using germ-free or gnotobiotic mice. The Gnoto / Axenic Facility of the Instituto Gulbenkian de Ciência hosted clients in previous user projects to perform on site e.g. histology, flow-cytometry or microscopy.